Identifying the region influencing the cis-mode of maturation of West Nile (Sarafend) virus using chimeric infectious clones

Li, J.; Bhuvanakantham, Raghavan; Howe, J.; Ng, Mah-Lee

doi:10.1016/j.bbrc.2005.06.150

Cited by 12 publications

(9 citation statements)

References 49 publications

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“…HEK293 cells were electroporated with 25 mg of RNAs synthesized from pWNS/mutant pWNS clones using RiboMAX RNA synthesis kit (Promega) as described earlier (Li et al ., ). The culture supernatants collected from the electroporated cells were passaged twice before used in downstream experiments.…”

Section: Methodsmentioning

confidence: 97%

“…In addition, QuikChange Site‐Directed Mutagenesis Kit (Stratagene) was used to mutate the required residues on WNV/DENV Myc‐tagged or HA‐tagged C protein. To introduce mutations in the full‐length infectious clones of WNV (Li et al ., ) and DENV (gift from Dr Andrew Davidson, Bristol University, UK), mutations were first introduced on a 1.3 kb carrier plasmid of WNV and DENV as described earlier (Li et al ., ). The 1.3 kb carrier plasmid contains 5′ UTR, C, prM and partial envelope sequences of flavivirus in pBR322 vector.…”

Section: Methodsmentioning

confidence: 99%

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West Nile virus and dengue virus capsid protein negates the antiviral activity of human Sec3 protein through the proteasome pathway

Bhuvanakantham

2013

Cell Microbiol

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Section: Methodsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

West Nile virus and dengue virus capsid protein negates the antiviral activity of human Sec3 protein through the proteasome pathway

Bhuvanakantham

2013

Cell Microbiol

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“…1. Modified pBR322 vector was generated as described earlier (Li et al, 2005a). Individual fragments corresponding to the four regions of the full-length genome were separately cloned into the modified vector and served as intermediate clones before sequential ligation into the vector to form pWNS.…”

Section: Methodsmentioning

confidence: 99%

The glycosylation site in the envelope protein of West Nile virus (Sarafend) plays an important role in replication and maturation processes

Bhuvanakantham

Howe

et al. 2006

Journal of General Virology

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The complete genome of West Nile (Sarafend) virus [WN(S)V] was sequenced. Phylogenetic trees utilizing the complete genomic sequence, capsid gene, envelope gene and NS5 gene/39 untranslated region of WN(S)V classified WN(S)V as a lineage II virus. A full-length infectious clone of WN(S)V with a point mutation in the glycosylation site of the envelope protein (pWNS-S154A) was constructed. Both growth kinetics and the mode of maturation were affected by this mutation. The titre of the pWNS-S154A virus was lower than the wild-type virus. This defect was corrected by the expression of wild-type envelope protein in trans. The pWNS-S154A virus matured intracellularly instead of at the plasma membrane as shown for the parental WN(S)V.

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“…It is known that WNV genome replication occurs in the cytoplasm in the perinuclear region and virus particles assemble and bud into the Endoplasmic Reticulum (ER) lumen. Subsequently virions are transported to the plasma membrane (PM) via the cellular secretory pathway to be released from cells by exocytosis [5-8]. …”

Section: Introductionmentioning

confidence: 99%

Identification of conserved motifs in the Westnile virus envelope essential for particle secretion

et al. 2013

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BackgroundEnveloped viruses utilize cellular membranes to bud from infected cells. The process of virion assembly and budding is often facilitated by the presence of certain conserved motifs within viral proteins in conjunction with cellular factors. We hence examined the West Nile Virus (WNV) Envelope protein for the presence of any such motifs and their functional characterization.ResultsWe identified conserved 461PXAP464 and 349YCYL352 motifs in the WNV envelope glycoprotein bearing resemblance to retroviral late domains. Disruptive mutations of PXAP to LAAL and of the highly conserved Cys350 in the YCYL motif, led to a severe reduction in WNV particle production. Similar motifs in case of retroviruses are known to interact with components of host sorting machinery like PXAP with Tsg101 and YXXL with Alix. However, in the case of WNV, siRNA mediated depletion of Alix or Tsg101 did not have an effect on WNV release. Molecular modeling suggested that while the 461PXAP464 motif is surface accessible and could potentially interact with cellular proteins required for WNV assembly, the 349YCYL352 motif was found to be internal with Cys350 important for protein folding via disulphide bonding.ConclusionsThe conserved 461PXAP464 and 349YCYL352 motifs in the WNV envelope are indispensable for WNV particle production. Although these motifs bear sequence similarity to retroviral late domains and are essential for WNV assembly, they are functionally distinct suggesting that they are not the typical late domain like motifs of retroviruses and may play a role other than Alix/Tsg101 utilization/dependence.

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Identifying the region influencing the cis-mode of maturation of West Nile (Sarafend) virus using chimeric infectious clones

Cited by 12 publications

References 49 publications

West Nile virus and dengue virus capsid protein negates the antiviral activity of human Sec3 protein through the proteasome pathway

West Nile virus and dengue virus capsid protein negates the antiviral activity of human Sec3 protein through the proteasome pathway

The glycosylation site in the envelope protein of West Nile virus (Sarafend) plays an important role in replication and maturation processes

Identification of conserved motifs in the Westnile virus envelope essential for particle secretion

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